KG-1 Whole Cell Lysate

SDS-Page: KG-1 Whole Cell Lysate [NBP3-05284] - Lane 1: KG-1 Whole Cell Lysate (10ug) Reduced. Lane 2: Opal Prestained Molecular Weight Markers Lane 3: KG-1 Whole Cell Lysate (10ug) Non-Reduced. Results show wide range of molecular weight bands with no signs of degradation.

• Reactivity: Hu
• Applications: WB, PAGE

KG-1 Whole Cell Lysate Summary

• Description: Store vial at -70C or COLDER. For extended storage, aliquot contents to minimize freeze/thaw cycles.
• Preparation Method: KG-1 cells were grown in Iscove's Modified Dulbecco's Medium supplemented with 20% fetal bovine serum. Cells were washed with PBS and then incubated on ice in modified RIPA buffer, containing 150 mM sodium chloride, 50 mM Tris HCl, pH 7.4, 1 mM EDTA, 1.0% NP-40, 0.5% sodium deoxycholic acid , 0.1% SDS and 0.01% (w/v) sodium azide to lyse the cells. Protein integrity was ensured using a cocktail of protease inhibitors with broad specificity for the inhibition of aspartic, cysteine, and serine proteases as well as aminopeptidases (0.1 mM AEBSF HCl, 0.08 uM Aprotinin, 5 uM Bestatin, 1.5 uM E-64, 2 uM Leupeptin Hemisulfate, 1 uM Pepstatin A). Phosphatase inhibitors 1 mM NaF and 1 mM Na3VO4 were also added. Cell debris was removed by centrifugation. Protein concentration was determined by a modified BCA assay using a commercially available kit. Protein concentration was adjusted to 1 mg/mL with 2X Sample buffer.
• Purity: Multi-step

Applications/Dilutions

• Dilutions:
  • SDS-Page
  • Western Blot
• Application Notes: This product has been tested by SDS-PAGE. Ready-to-use lysates are especially prepared as positive controls for separation by SDS-PAGE and subsequent western blot analysis. Lysates are prepared in denaturing buffer WITHOUT dissociating agents (i.e. no 2-mercaptoethanol or dithiothreitol has been added). Heat lysate to 95C for 5 minutes and rapidly cool. If dissociating conditions are desired, add reducing agent prior to heating. The recommended loading volume per lane is 10-20 uL depending on the size format of your gel.

Packaging, Storage & Formulations

• Storage: Store at -70C. Avoid freeze-thaw cycles.
• Buffer: 1X SDS-PAGE Sample Buffer (62.5 mM Tris HCl, 2% SDS, 10% (v/v) Glycerol and 0.005% bromophenol blue, pH 6.8)
• Preservative: No Preservative
• Purity: Multi-step

Lysate Details for Array

• Type: Cell
• Subcellular Fraction: Whole

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.